口服食用燕窝后小鼠血清和组织中游离和共轭硅酸、Neu5Ac、Neu5Gc 和 KDN 水平的变化:LC-MS/MS 定量分析

Meng-Hua Wang, Zhi-Fan Wang, Man Yuan, Chun-Guo Yang, Dong-Liang Wang, Shu-Qi Wang
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引用次数: 0

摘要

食用燕窝具有多种生物活性,其中的主要成分是硅酸。硫辛酸是神经氨酸的一组九碳 N-乙酰化衍生物,在 C2 位含有一个酮基,在许多生物过程中发挥着重要作用。为了验证口服燕窝是否会改变体内硅酸成分的含量和分布,研究人员开发了一种液相色谱-质谱法,用于定量分析血清和组织中的硅酸含量。在负离子模式下,流动相为 0.1% 甲酸水溶液(A)和乙腈(v/v)(B)。以 60% 的 B 进行等度洗脱,时间为 0-15 分钟。色谱分离采用 Morphling HILIC Amide 色谱柱(2.1 mm × 150 mm,5 μm),流速为 0.5 mL min-1。结果表明,典型校正曲线的相关系数均大于 0.995,表现出良好的线性关系。在小鼠口服燕窝七天后,同时检测血清和不同组织中游离型和共轭型 N-乙酰神经氨酸(Neu5Gc)、N-乙酰神经氨酸(Neu5Ac)和 2-酮基-3-脱氧-D-甘油-D-半乳糖醛酸(KDN)的含量。我们的研究发现,口服燕窝能显著提高小鼠血清和脾肺组织中总硅酸(Neu5Gc + Neu5Ac + KDN)的浓度,这可能与燕窝的抗炎和免疫功能有关,但还需要进一步的研究来验证。在脑组织中,Neu5Ac是最主要的序列酸;在血清和其他组织(包括心、肝、脾、肺和肾)中,Neu5Gc是最主要的序列酸。此外,我们还发现除肝脏组织外,其他组织中的 Neu5Ac 和 Neu5Gc 均以共轭形式为主。总之,所建立的方法线性关系好、准确度高,能有效地分离血清或组织中的基质和内源性物质,能有效地检测血清和组织中游离型和共轭型硅烷酸的分布和浓度,有利于燕窝和硅烷酸的研究和开发利用。
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Changes in the Serum and Tissue Levels of Free and Conjugated Sialic Acids, Neu5Ac, Neu5Gc, and KDN in Mice after the Oral Administration of Edible Bird’s Nests: An LC–MS/MS Quantitative Analysis
Edible bird’s nests have a variety of biological activities, the main components of which are sialic acids. Sialic acids are a group of nine-carbon N-acetylated derivatives of neuraminic acid containing a keto group at position C2 and play important roles in many biological processes. To verify whether the oral administration of edible bird’s nests would change the content and distribution of sialic acid components in vivo, a liquid chromatography–mass spectrometry method for the quantitative analysis of sialic acid levels in serum and tissues was developed. In the negative ion mode, the mobile phases consist of 0.1% formic acid in water (A) and acetonitrile (v/v) (B). Isocratic elution was performed with 60% B for 0−15 min. The chromatographic separation was performed on a Morphling HILIC Amide column (2.1 mm × 150 mm, 5 μm) at a flow rate of 0.5 mL min−1. The results showed that the correlation coefficients of the typical calibration curves were all higher than 0.995, exhibiting good linearity. The levels of free and conjugated forms of N-glycolylneuraminic acid (Neu5Gc), N-acetylneuraminic acid (Neu5Ac), and 2-keto-3-deoxy-D-glycero-D-galactonononic acid (KDN) in the serum and different tissues were simultaneously detected after the oral administration of the edible bird’s nests at a daily dose of 300 and 700 mg Kg−1 for seven days in mice. Our study found that the oral administration of edible bird’s nests can significantly increase the concentration of total sialic acids (Neu5Gc + Neu5Ac + KDN) in serum and spleen and lungs tissues, which may be related to the anti-inflammatory and immune function of edible bird’s nest, but further studies are needed to verify this. Neu5Ac was the dominant sialic acid in brain tissue, and Neu5Gc was the dominant sialic acid in serum and other tissues, including heart, liver, spleen, lungs, and kidney. Moreover, we found that the forms of Neu5Ac and Neu5Gc were mainly conjugated in all groups except liver tissue. In conclusion, the method we established had good linearity and accuracy; it allowed the analytes to be effectively separated from the matrix and endogenous substances in serum or tissues, so it could effectively detect the distribution and concentration of free and conjugated forms of sialic acids in serum and tissues, which was beneficial to the research and exploitation of edible bird’s nests and sialic acids.
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